Our study aimed to create a more effective model for researching adrenocortical diseases by improving the differentiation process of human induced pluripotent stem cells (hiPSCs) into adrenocortical cells. We wanted a system that mimics natural hormone production and allows for easy genetic manipulation. By expressing a key factor, steroidogenic factor-1 (SF-1), and adding essential niche factors, we successfully generated cells that produce adrenal steroids like aldosterone. These cells showed gene expression patterns similar to those of fetal and adult adrenal cortex cells, responded to natural stimuli, and kept their ability to proliferate. By using CRISPR/Cas9 to introduce a KCNJ5 mutation associated with primary aldosteronism, we observed increased aldosterone production and cell growth, which are crucial for studying tumor development. This new system offers a valuable tool for better understanding adrenocortical diseases and developing new treatments.Here, we provide our annotated single-cell RNA sequencing data as a Seurat object (Seurat v5.0.1).